Trichothecene producers, detection

D. Other sequence sources for detection of trichothecene producers... 

Various sequence sources other than genes from the trichothecene biosynthesis pathway were used to set up PCR-based systems for identification and detection of trichothecene producers. Highly specific... 

Agodi, A., Barchitta, M., Ferrante, M., Sciacca, S., and Niessen, L. (2005). Detection of trichothecene producing Fusarium spp. by PCR Adaptation, validation and application to fast food. Italian J. Public Health 3, 7-11. 

Niessen, M. L., and Vogel, R. F. (1998). Group specific PCR-detection of potential trichothecene-producing Fusarium-spedes in pure cultures and cereal samples. Syst. Appl. Microbiol. 21, 618-631. 

Niessen, L., Schmidt, H., and Vogel, R. F. (2004). The use of triS gene-sequences for PCR detection and taxonomy of trichothecene-producing species in the Fusarium section Sporotrichiella. Int. J. Food Microbiol. 95, 305-319. 

Trichothecene mycotoxins are highly irritating to the skin and mucous membranes in all species. Skin irritation was experienced by laboratory workers accidentally exposed to trichothecene producing fungal cultures or crude extracts of T-2 toxin. Skin irritation has also been used as a basis for a semiquantitative bioassay to detect these toxins (Bamburg... 

Another important application arising from the work with trichodiene synthase has been the development of Tri5 specific polymerase chain reaction-based assays for the detection of potential trichothecene-producing Fusarium species in pure culture and in contaminated grain (269,275,276). 

Niessen ML, Vogel RE (1998) Group Specific PCR-Detection of Potential Trichothecene-Producing Fusarium Species in Pure Cultures and Cereal Samples. System Appl Microbiol 21 618... 

Nicholson P, Simpson D R, Wilson A H, Chandler E and Thomsett M (2004), Detection and differentiation of trichothecene enniatin-producing Fusarium species on small-grain cereals , Europ. J. Plant Pathol., 110, 503-514. 

Bluhm, B. H., Flaherty, J. E., Cousin, M. A., and Woloshuk, C. P. (2002). Multiplex Polymerase Chain Reaction assay for the differential detection of trichothecene- and fumonisin-producing species of Fusarium in cornmeal. /. Food Prot. 65,1955-1961. 

Kristensen, R., Berdal, K. G., and Holst-Jensen, A. (2006a). Simultaneous detection and identification of trichothecene- and moniliformin-producing Fusarium species based on multiplex SNP analysis. /. Appl. Microbiol, pp. 11. doi 10.1111/j.l365-2672.2006.03166.x. 

In our research, we proposed a post amplification analytical method to detect F. culmorum, a pathogen causing foot rot and head blight diseases in cereals, and can produce mycotoxins such as zearalenone, deoxynivalenol, and other trichothecenes that can enter the food chain. The early identification of this fungal pathogen is therefore recommended in order to avoid crop losses and protect consumer health... 

For the biosynthesis of a model type A trichothecene T-2 toxin, all the necessary Tri genes were already identified.226 However, there still remain two Tri genes as-yet-unidentified for the biosynthesis of 4-ANIV, a model type B trichothecene. These include genes encoding DHC C-8 oxidoreductase, whose activity was also detected from F. verticittioides (teleomorph genus Gibberella), which does not produce trichothecenes,274 and trichothecene C-15 deacetylase,292 whose distribution and substrate specificity are not well examined. 

To overcome the difficulties encountered with the bioassays and TLC methods, immunoassays using specific polyclonal and monoclonal antibodies have been developed for most of the major trichothecene mycotoxins and their metabolites.73 These antibodies have been used to produce simple, sensitive, and specific radioimmunoassays (RIAs) and enzyme-linked immunosorbent assays (ELISAs) for the mycotoxins. In the presence of the sample matrix, the lower detection limits for identification of trichothecene mycotoxins by RIA is about 2 to 5 ppb73 and by ELISA, 1 ppb.74 We conclude that immunoassays are useful tools for screening biomedical samples for evidence of a biological warfare attack with trichothecene mycotoxins. 

To follow the contamination of grains with mycotoxins in primary production in Slovenia, altogether 175 samples of cereals produced in Slovenia were collected at farms in the years 2007, 2008, and 2009. Among them, there were 79 samples of maize, 39 samples of barley, and 34 samples of wheat. In the collected samples, mycotoxins were determined. For the determination of trichothecenes, the analytical procedure mentioned above was used. As positive, samples with the concentration of mycotoxins higher than limit of detection (LOD) were designated. 

Deoxynivalenol (DON) is the most important trichothecene worldwide and is often detected in small cereal grains such as barley, oats and wheat. Due to relatively good thermal stability DON can be transmitted from contaminated barley into the final product (Schwarz et al., 1995). DON is frequently detected in barley and in commercial beer (discussed in Section 6.6.2). The occurrence of DON is largely dependent on weather conditions in the particular location and year. DON is predominantly produced by F. culmorum and F. graminearum species. 

GC/FTIR data have also contributed to the structure elucidation of other compounds of biological origin, such as mycotoxins, which are formed by fungal activity in food products under specific environmental conditions of moisture, temperature, and host. Trichothecene mycotoxins, secondary fungal metabolites produced by species of mold, are a natural contaminant of feedstuffs and food. Because they can be toxic to humans and animals, their detection is important. Sehat et al., utilized GC/MI-FTIR and GC/MS to analyze grains for these contaminants. 

---